Chitosan/HPMC-based mucoadhesive film co-loaded with fluconazole and ofloxacin for management of polymicrobial keratitis.

Dynamic emergence of microbial keratitis (MK) requires a promising therapeutic arsenal of antifungal and antibacterial agents like voriconazole (VCZ) and moxifloxacin (MOXI), respectively. Parallelly, another paradigm of MK associated with ulcerative wounds cannot be left unnoticed and requires antifibrotic remedy (pirfenidone, PIR) as an authalic antimicrobial to retain the primordial vision. For designing an effective clinical cure, a combination of these three agents is required at a therapeutic dosage regimen. Following the quest, we have developed a simple and sensitive LC-MS/MS bioanalytical method for simultaneous quantification of VCZ, MOXI, and PIR in rabbit lacrimal fluid. The method was validated as per US-FDA norms using ketoconazole as an internal standard for linearity, accuracy-precision, matrix effect, dilution integrity, selectivity, and stability. The five minutes chromatographic set-up includes isocratic elution with a C18 column using MeOH (80%, v/v) and ultrapure water containing 0.2% formic acid (20%, v/v), respectively. The MS-based analyte detection was achieved in ESI+ multiple reaction monitoring mode. The sample extraction was performed using the protein precipitation method with minimal sample size. The validated methodology was employed to determine the ocular pharmacokinetics profile of marketed formulations containing VCZ, MOXI, and PIR in rabbit lacrimal matrix.

Simultaneous estimation of voriconazole, moxifloxacin, and pirfenidone in rabbit lacrimal matrix using LC-MS/MS: an application to preclinical ocular pharmacokinetics.

Fenugreek seeds are used in numerous marketed herbal formulations having therapeutic benefits. Some of its bioactive components including 4-hydroxyisoleucine (4-HIL), trigonelline (TG), raffinose (RFO) and pinitol (PT) are reported to have potential therapeutic activity such as antibacterial, antidiabetic, stomach stimulant, anti-invasive, hyperandrogenism and other allied diseases including polycystic ovary syndrome (PCOS). A fully validated, selective and sensitive bioanalytical method for simultaneous rapid quantification of the above mentioned bioactive components has been developed using hyphenated liquid chromatography electrospray tandem mass spectrometry. Separation of the analytes was achieved within 5 minutes by gradient elution in C18 column at 0.5 mL/min flow rate. Plasma protein precipitation technique was employed to isolate the analytes from the samples. Oral pharmacokinetic profile of the four bioactive components in SD rats was then further evaluated using non compartmental analysis using Phoenix WinNonlin software.

Simultaneous pharmacokinetic assessment of phytopharmaceuticals from Fenugreek extract by liquid chromatography-tandem mass spectrometry in Sprague Dawley rats.

A novel, quick, and precise RP-UHPLC analytical method for simultaneous determination of moxifloxacin (MFX), voriconazole (VCZ), and pirfenidone (PIR) was developed and validated according to the ICH guidelines using QbD driven response surface Box Behnken design. The developed method was validated considering selectivity, sensitivity, linearity, accuracy-precision, robustness, stability, the limit of detection and limit of quantification, respectively. The resolution between MFX, VCZ, and PIR was achieved using gradient elution protocol against Waters Symmetry Shield C18 column (150 × 4.6 mm2 , 5 μ) using Agilent 1290, Infinity II series LC system. The method was applied to quantitatively estimate proprietary and in-house prepared pharmaceutical topical ophthalmic formulations containing MFX, VCZ, and PIR at λmax of 296, 260, and 316 nm. The method is sensitive enough to detect upto 0.1 ppm of analytes in the formulation. The method was further exploited to study and identify the possible degradation products of the analytes. The proposed chromatographic method is simple, economical, reliable, and reproducible. In conclusion, the developed method could be applicable for routine quality control analysis of single or a combination of MFX, VCZ, and PIR-containing unit or bulk dosage forms in pharmaceutical industries and research organizations working on drug discovery and drug development.

A QbD-led Simple and Sensitive RP-UHPLC method for Simultaneous Determination of Moxifloxacin, Voriconazole, and Pirfenidone: An Application to Pharmaceutical Analysis.

LC-MS/MS based quantification of steroidal biomarkers in polycystic ovary syndrome induced rats.

Simultaneous determination of fluconazole and ofloxacin in rabbit tear fluid by LC-MS/MS: Application to ocular pharmacokinetic studies.

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Simultaneous determination of fluconazole and ofloxacin in rabbit tear fluid by LC-MS/MS: Application to ocular pharmacokinetic studies.